• Title/Summary/Keyword: amylolytic

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Xanthomonas euvesicatoria Causes Bacterial Spot Disease on Pepper Plant in Korea

  • Kyeon, Min-Seong;Son, Soo-Hyeong;Noh, Young-Hee;Kim, Yong-Eon;Lee, Hyok-In;Cha, Jae-Soon
    • The Plant Pathology Journal
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    • v.32 no.5
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    • pp.431-440
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    • 2016
  • In 2004, bacterial spot-causing xanthomonads (BSX) were reclassified into 4 species-Xanthomonas euvesicatoria, X. vesicatoria, X. perforans, and X. gardneri. Bacterial spot disease on pepper plant in Korea is known to be caused by both X. axonopodis pv. vesicatoria and X. vesicatoria. Here, we reidentified the pathogen causing bacterial spots on pepper plant based on the new classification. Accordingly, 72 pathogenic isolates were obtained from the lesions on pepper plants at 42 different locations. All isolates were negative for pectolytic activity. Five isolates were positive for amylolytic activity. All of the Korean pepper isolates had a 32 kDa-protein unique to X. euvesicatoria and had the same band pattern of the rpoB gene as that of X. euvesicatoria and X. perforans as indicated by PCR-restriction fragment length polymorphism analysis. A phylogenetic tree of 16S rDNA sequences showed that all of the Korean pepper plant isolates fit into the same group as did all the reference strains of X. euvesicatoria and X. perforans. A phylogenetic tree of the nucleotide sequences of 3 housekeeping genes-gapA, gyrB, and lepA showed that all of the Korean pepper plant isolates fit into the same group as did all of the references strains of X. euvesicatoria. Based on the phenotypic and genotypic characteristics, we identified the pathogen as X. euvesicatoria. Neither X. vesicatoria, the known pathogen of pepper bacterial spot, nor X. perforans, the known pathogen of tomato plant, was isolated. Thus, we suggest that the pathogen causing bacterial spot disease of pepper plants in Korea is X. euvesicatoria.

Evaluation and Genome Mining of Bacillus stercoris Isolate B.PNR1 as Potential Agent for Fusarium Wilt Control and Growth Promotion of Tomato

  • Rattana Pengproh;Thanwanit Thanyasiriwat;Kusavadee Sangdee;Juthaporn Saengprajak;Praphat Kawicha;Aphidech Sangdee
    • The Plant Pathology Journal
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    • v.39 no.5
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    • pp.430-448
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    • 2023
  • Recently, strategies for controlling Fusarium oxysporum f. sp. lycopersici (Fol), the causal agent of Fusarium wilt of tomato, focus on using effective biocontrol agents. In this study, an analysis of the biocontrol and plant growth promoting (PGP) attributes of 11 isolates of loamy soil Bacillus spp. has been conducted. Among them, the isolates B.PNR1 and B.PNR2 inhibited the mycelial growth of Fol by inducing abnormal fungal cell wall structures and cell wall collapse. Moreover, broad-spectrum activity against four other plant pathogenic fungi, F. oxysporum f. sp. cubense race 1 (Foc), Sclerotium rolfsii, Colletotrichum musae, and C. gloeosporioides were noted for these isolates. These two Bacillus isolates produced indole acetic acid, phosphate solubilization enzymes, and amylolytic and cellulolytic enzymes. In the pot experiment, the culture filtrate from B.PNR1 showed greater inhibition of the fungal pathogens and significantly promoted the growth of tomato plants more than those of the other treatments. Isolate B.PNR1, the best biocontrol and PGP, was identified as Bacillus stercoris by its 16S rRNA gene sequence and whole genome sequencing analysis (WGS). The WGS, through genome mining, confirmed that the B.PNR1 genome contained genes/gene cluster of a nonribosomal peptide synthetase/polyketide synthase, such as fengycin, surfactin, bacillaene, subtilosin A, bacilysin, and bacillibactin, which are involved in antagonistic and PGP activities. Therefore, our finding demonstrates the effectiveness of B. stercoris strain B.PNR1 as an antagonist and for plant growth promotion, highlighting the use of this microorganism as a biocontrol agent against the Fusarium wilt pathogen and PGP abilities in tomatoes.

Isolation and Characteristics of Bacteriocin-producing Bacteria from the Intestine of Duck for Probiotics (오리로부터 박테리오신을 생산하는 프로바이오틱 미생물의 분리 및 특성)

  • Shin, M.S.;Han, S.K.;Ji, A.R.;Ham, M.R.;Kim, K.S.;Lee, W.K.
    • Journal of Animal Science and Technology
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    • v.49 no.5
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    • pp.621-632
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    • 2007
  • The aim of this study was to isolate and characterize bacteriocin-producing bacteria from the intestine of duck to use as probiotics for livestock. A total of 416 strains were isolated from the small intestine and cecum of ducks and 13 isolates were finally selected after determinging inhibitory activity against pathogenic indicators by spot-on-lawn method. The selected strains were identified as Lactobacillus salivarius JWS 58, Lactobacillus plantarum JWS 1354, Pediococcus pentosaceus JWS 939, 7 strains of enterococci, and 3 strains of Escherichia coli. Lact. salivarius JWS 58, Ent. faecium JWS 833, and Ped. pentosaceus JWS 939 showed a strong inhibitory activity against Listeria monocytogenes. E. coli JWS 108 inhibited the growth of E. coli and Staphylococcus aureus. Lact. salivarius JWS 58 strain survived almost 50% in pH 2.5 phosphate buffer for 2 hr. Ped. pentosaceus JWS 939 and Lact. plantarum JWS 1354 showed strong amylolytic activity. These results suggest that a combination of bacteriocins or multispecies probiotics of the selected strains has a strong potential of alternative to antibiotics in livestock production.

Effects of Eucalyptus Crude Oils Supplementation on Rumen Fermentation, Microorganism and Nutrient Digestibility in Swamp Buffaloes

  • Thao, N.T.;Wanapat, M.;Cherdthong, A.;Kang, S.
    • Asian-Australasian Journal of Animal Sciences
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    • v.27 no.1
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    • pp.46-54
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    • 2014
  • This study was conducted to investigate the effects of eucalyptus (E. Camaldulensis) crude oils (EuO) supplementation on voluntary feed intake and rumen fermentation characteristics in swamp buffaloes. Four rumen fistulated swamp buffaloes, body weight (BW) of $420{\pm}15.0$ kg, were randomly assigned according to a $2{\times}2$ factorial arrangement in a $4{\times}4$ Latin square design. The dietary treatments were untreated rice straw (RS) without EuO (T1) and with EuO (T2) supplementation, and 3% urea-treated rice straw (UTRS) without EuO (T3) and with EuO (T4) supplementation. The EuO was supplemented at 2 mL/h/d in respective treatment. Experimental animals were kept in individual pens and concentrate mixture was offered at 3 g/kg BW while roughage was fed ad libitum. Total dry matter and roughage intake, and apparent digestibilites of organic matter and neutral detergent fiber were improved (p<0.01) by UTRS. There was no effect of EuO supplementation on feed intake and nutrient digestibility. Ruminal pH and temperature were not (p>0.05) affected by either roughage sources or EuO supplementation. However, buffaloes fed UTRS had higher ruminal ammonia nitrogen and blood urea nitrogen as compared with RS. Total volatile fatty acid and butyrate proportion were similar among treatments, whereas acetate was decreased and propionate molar proportion was increased by EuO supplementation. Feeding UTRS resulted in lower acetate and higher propionate concentration compared to RS. Moreover, supplementation of EuO reduced methane production especially in UTRS treatment. Protozoa populations were reduced by EuO supplementation while fungi zoospores remained the same. Total, amylolytic and cellulolytic bacterial populations were increased (p<0.01) by UTRS; However, EuO supplementation did not affect viable bacteria. Nitrogen intake and in feces were found higher in buffaloes fed UTRS. A positive nitrogen balance (absorption and retention) was in buffaloes fed UTRS. Supplementation of EuO did not affect nitrogen utilization. Both allantoin excretion and absorption and microbial nitrogen supply were increased by UTRS whereas efficiency of microbial protein synthesis was similar in all treatments. Findings of present study suggested that EuO could be used as a feed additive to modify the rumen fermentation in reducing methane production both in RS and UTRS. Feeding UTRS could improve feed intake and efficiency of rumen fermentation in swamp buffaloes. However, more research is warranted to determine the effect of EuO supplementation in production animals.

The Effect of Nutritional Balance between Carbon and other Nutrient Sources on the Growth of Sporobolomyces holsaticus (탄소원과 다른 영양원간의 영양균형이 Sporobolomyces holsaticus의 균체생육도에 미치는 영향)

  • Park, Wan-Soo;Koo, Young-Jo;Shin, Dong-Hwa;Min, Byung-Yong
    • Korean Journal of Food Science and Technology
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    • v.15 no.1
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    • pp.56-61
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    • 1983
  • Direct production of biomass from starch using amylolytic yeast, Sporobolomyces holsaticus FRI Y-5 was studied with varying the ratios between carbon and other nutrient sources in the medium. It was investigated under condition of constant C/P and C/S ratio to influence the initial concentration of starch $(S_o)$ and C/N ratio on its growth which is described as the specific growth rate $({\mu})$, cell yield (Y), the maximum concentration of cell $(X_m)$, and productivity (P). They were very dependent on both $S_o$ and C/N ratio. The form of the relationship between and ${\mu}$ and $S_o$ was observed to be similar to saturation kinetics at C/N = 100 but presented substrate inhibition at other C/N ratios. As $S_o$ was changed from 22.5 to 90 g/l, Y was observed to vary with C/N ratios but seemed to decrease as a wholes. $X_m$ was linearly related to $S_o$ at more than C/N = 50 but at less than C/N = 10 substrate inhibition was presented. P increased suddenly to $S_o$ = 45 g/l and then changed decreasingly at less than C/N = 50, but at more than C/N = 100 it changed increasingly. The effect of C/P ratio and C/S ratio on the yeast growth was also investigated at constant $S_o$ and C/N ratio. ${\mu}$ was dependent on C/P and C/S ratios, but Y, independent on them. But $X_m$ was reliant upon C/P ratio but not upon C/S ratio.

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Preparation and Utilization of High-Protein Rice Flour (고단백 쌀가루의 제조 및 이용)

  • Cha, Jae-Ho;Kwon, Ki-Hwa;Park, Kwan-Hwa;Chang, Hak-Gil
    • Korean Journal of Food Science and Technology
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    • v.20 no.6
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    • pp.840-844
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    • 1988
  • High-protein rice flour (HPRF) was prepared by an enzymatic process using ${\alpha}-amylase$ and g1ucoamylase without cooking Process and the feasibility of HPRF as infants foods was tested. Rice flour slurry was treated with 0.25% ${\alpha}-amylase$ and 0.5% glucoamylase at $55^{\circ}C$ for 24hrs. After saccharification, the digested rice slurry was centrifuged and the precipitated paste, was then heat-dried to obtain HPRF. The protein content of the HPRF was 20.8%. On the other hand, the supernatant of glucose enriched solution was decolourized, deionized and then isomerised to furctose at $60^{\circ}C$ for 100min by using immobilized glucose isomerase column. The high-fructose solution (HFS) contained 56% glucose, 42% fructose and 2% oligosaccharide. The nutritional quality of the HPRF was compared with milk protein and soybean protein in weight gain, feed efficiency ratio (FER), protein efficiency ratio (PER) and liver weight. HPRF was almost the same in all items with milk and soybean protein, but significantly superior to rice flour group.

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Cloning of Isoamylase Gene of Pectobacterium carotovorum subsp. carotovorum LY34 and Identification of Essential Residues of Enzyme (Pectobacterium carotovorum subsp. carotovorum LY34에서 Lsoamylase 유전자 클로닝 및 효소 활성의 필수 잔기 확인)

  • Cho, Kye-Man;Kim, Eun-Ju;Math, Renukaradhya K.;Asraful Islam, Shah Md.;Hong, Sun-Joo;Kim, Jong-Ok;Shin, Ki-Jae;Lee, Young-Han;Kim, Hoon;Yun, Han-Dae
    • Journal of Life Science
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    • v.17 no.9 s.89
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    • pp.1182-1190
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    • 2007
  • The gene encoding for isoamylase of the Pectobacterium carotovorum subsp. carotovorum (Pcc) LY34 was cloned and expressed into Escherichia coli $DH5{\alpha}$. Isoamylase catalyzes the hydrolysis of ${\alpha}-1,6-glycosidic$ linkages specifically in amylopectin, glycogen, and derived oligosaccharides, while the enzyme did not hydrolyze ${\alpha}-1,4-glycosidic$ linkages of amylose. The isoamylase gene (glgX) had an open reading frame of 1,977 bp encoding 658 amino acid residues with a calculated molecular weight of 74,188 Da. The molecular weight of the enzyme was also estimated to be 74 kDa by activity staining of a SDS-PA gel. The mature GlgX had a calculated pI of 4.91. Isoamylase from Pcc LY34 had 70% amino acid identity with isoamylase from Pectobacterium chrysanthemi and contained the four regions conserved among all amylolytic enzymes. The isoamylase was optimally active at pH 7.0 and $40^{\circ}C$. GlgX was $Ca^{2+}-dependent$. The changes of Asp-335, Glu-370, and Asp-442 into Ala, respectively, using site-directed mutagenesis techniques showed that three residues are essential to isolamyalse (GlgX) activity. The sequences around those residues were highly conserved in isoamylase of different origins and GlgX of the glg operon in glycongen biosynthesis.

Production of L-Lactic Acid from Soluble Starch by Enterococcus sp. JA-27. (Enterococcus sp. JA-27에 의한 가용성 전분으로부터 L형 젖산의 생산)

  • 김경아;김미경;장경린;전홍기
    • Microbiology and Biotechnology Letters
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    • v.31 no.3
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    • pp.250-256
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    • 2003
  • Lactic acid bacteria with amylolytic and acid producing activities can ferment starch directly to lactic acid thereby producing a monomer for the production of biodegradable poly lactic acid (PLA). In this study, the strain producing L-lactic acid from soluble starch was isolated from Nuruk. The isolated strain was identified as Enterococcus sp. through its morphological, cultural, biochemical characteristics as well as the 16S rDNA sequence analysis, and named Enterococcus sp. JA-27. Enterococcus sp. JA-27 produced exclusively L-lactic acid from soluble starch as a carbon source. The optimal conditions for the maximum production of L-lactic acid from Enterococcus sp. JA-27 were 30 C, pH 8, 1.5 % soluble starch as a substrate and 3.5 % tryptone as a nitrogen source, 0.1 % $K_2$$HPO_4$, 0.04 % $MgSO_4$. $7H_2$O, 0.014 % $MnSO_4$$.$4$H_2O$, 0.004% $FeSO_4$$.$$7H_2$O. Batch and fed batch culture were carried out and the former was more effective. L-Lactic acid production in the optimum medium was significantly increased in a 7 L jar fermenter, where the maximum L-lactic acid concentration was 3 g/L. For the purification of lactic acid in fermented broth, two stage ionexchange column chromatographies were employed and finally identified by HPLC.

A Study on the Relationships Between the Epiphytic Microbes and the Blight of Porphyra Species from the Coastal Waters of the Yellow Sea, Korea -II. Seasonal Variation of Heterotrophic Bacteria in the Surrounding Seawater of Porphyra Farming Area Near Mokp'o- (서해산 김 엽체상의 미소생물과 김의 병해와의 관계에 대한 연구 -II. 목포 인근 김 양식장 주변해수에서의 종속영양세균의 계절적 변화-)

  • KIM Jeong-Hee;LEE Geon-Hyoung;SHIN Yoon-Keun;KIM Joong-Rae
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.25 no.4
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    • pp.307-313
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    • 1992
  • To study the seasonal variation of heterotrophic bacteria near the surrounding seawater of Porphyra forming area, samples were collected in the intertidal waters of Mokp'o of the Yellow Sea from February to December, 1990. Annual distribution of heterotrophic bacteria ranged from $7.5\times10^2\;to\;1.1\times10^5\;cfu/ml,$ annual distribution of physiological characteristic bacteria ranged from $5.0\times10\;to\;4.34\times10^4\;cfu/ml$ for proteolytic bacteria, from 0 to $1.35\times10^4\;cfu/ml$ for lipolytic bacteria and from 0 to $1.2\times10^4\;cfu/ml$ for amylolytic bacteria. Sixty-five percent of isolates from seawater were rods, and $76.7\%$ of isolates were Gram-negative. Most isolates were mesophiles and showed utilization of various carbon sources such as glucose, maltose, lactose, sucrose and arabinose. Most isolates also showed tolerance to a broad range of salt concentration. Dominant genus in seawater were Flavobacterium spp., in February, Moraxella spp., Acinetobacter spp. in March, Bacillus spp., Chromobacterium spp., Micrococcus spp., Vibrio spp. in July and Chromobacterium spp., Micrococcus spp., Bacillus spp. in November.

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Fermentation and Quality Characteristics of Cheonggukjang with Addition of Dropwort (Oenanthe javanica D.C.) Powder (미나리 분말을 첨가한 청국장의 발효 및 품질특성)

  • Lee, Shin-Ho;Kim, Jin-Hak
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.42 no.7
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    • pp.1133-1138
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    • 2013
  • Characteristics of Cheonggukjang with addition of different dropwort (Oenanthe javanica D.C.) powders were investigated. The selected strain, with proteolytic, amylolytic, and antimicrobial activity, was identified as B. subtilis RS-9, using 16S rRNA analysis. The Cheonggukjang was prepared with cooked soybean without dropwort (Control), 0.5% raw dropwort powder (DW0.5), and 1% raw dropwort powder (DW1), 0.5% steamed dropwort powder (SDW0.5), and 1% steamed dropwort powder (SDW1) were added, respectively. The changes in pH of Cheonggukjangs with addition of dropwort powder were lower than those of control during fermentation for 72 hr at $40^{\circ}C$. The total aerobes of the various Cheonggukjangs reached 8.88 (control), 8.82 (DW0.5), 8.70 (DW1), 8.85 (SDW0.5), and 8.75 (SDW1) log CFU/mL after fermentation for 72 hr at $40^{\circ}C$, respectively. The amino nitrogen and viscous substance contents of different dropwort powders added to Cheonggukangs were lower than those of control. The total polyphenol contents and ABTS radical scavenging ability of various Cheonggukjangs were increased by addition of dropwort powder and fermentation. The polyphenol contents and ABTS radical scavenging ability of SDW1 were $590.24{\mu}g/mL$ and 82.16% and showed the highest value among tested Cheonggukangs. The sensory quality of DW0.5 was higher in taste and overall acceptability, compared with other groups.